Sample preparation apparatus, sample preparation system, sample preparation method, and particle analyzer

ABSTRACT

The sample preparation apparatus includes a measurement unit configured to measure a sample containing particles acquired from a sample container and detect measurement target particles in the sample, a sample preparation unit capable of adjusting the concentration of measurement target particles in a sample acquired from a sample container and configured to prepare a measurement sample by mixing a sample and any of a plurality of types of particle detection reagents including a particle labeling substance, and a control unit for controlling the sample preparation unit so as to generate concentration information of the measurement target particles in the sample in the sample container based on the measurement data of the measurement unit and adjust the concentration of the measurement target particles in the sample acquired from the sample container according to the generated concentration information and the type of particle detection reagent used for preparing the measurement sample.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/992,524, filed on May 30, 2018, which claims priority from priorJapanese Patent Application No. 2017-108841, filed on May 31, 2017, theentire contents of which are each incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to a sample preparation apparatus, asample preparation system, a sample preparation method, and a particleanalyzer.

BACKGROUND

Japanese Patent Publication No. 07-301586 discloses a sample processingapparatus provided with concentration measuring means for measuring theconcentration of particle components contained in a liquid sample, and aconcentration unit for concentrating the sample by the filtering actionof a filter. WIPO Patent Publication No. 2009/122999 discloses a samplepreparation apparatus which generates concentration informationreflecting the concentration of epithelial cells contained in abiological sample when detecting cancerous cells among the epithelialcells contained in the biological sample, and controls the amount of thebiological sample to be supplied to a sample preparation unit based onthe concentration information.

The sample processing apparatus described in Japanese Patent PublicationNo. 07-301586 discloses concentrating the cells in the sample for thepurpose of improving the high reproducibility (probability error) of themeasurement result when the particle concentration is low in a particleanalyzer such as a flow cytometer. The sample preparation apparatusdescribed in WIPO Patent Publication No. 2009/122999 discloses adjustingan amount of a biological sample which is mixed with a staining liquidbased on the concentration of the epithelial cells in the biologicalsample obtained in a preliminary measurement so that the epithelialcells are appropriately stained with the staining liquid.

SUMMARY OF THE INVENTION

The sample processing apparatus described in Japanese Patent PublicationNo. 07-301586 and the sample preparation apparatus described in WIPOPatent Publication No. 2009/122999 both stain cell nuclei by simplestaining using pigments such as ethidium bromide, acridine orange,propidium iodide and the like, and detect the stained cells by anoptical detection method.

On the other hand, in current flow cytometric examinations, the type ofantigen measured by one examination request ranges from 10 to 30 typesper sample in order to diagnose one disease. For this reason,multi-color flow cytometry, in which several to ten kinds of antigensare simultaneously measured in a single analysis, is the main flowcytometry employed in flow cytometry. In order to detect all antigensthat have been requested to be examined, multicolor flow cytometricanalysis using several kinds of antibodies per analysis also needs to beperformed multiple times for each sample. The abundance of each antigenpresent in one cell also differs from antigen to antigen. In order toaccurately detect a target antigen, it is necessary to use afluorescently labeled antibody in an amount appropriate for eachantigen. Since stem cells such as hematopoietic stem cells have lowabundance ratios themselves in the living body, the amount of sampleitself to be analyzed has to be increased in order to reliably detectstem cells.

At present, the determination of the mixing ratio of a sample anddetection reagent and the determination of the necessary amount ofsample are carried out by the examiner in some form by obtaininginformation on the concentration of particles in the sample. Allpreparations of measurement samples based on the determined contents arealso performed by the examiner. From this situation, accurate flowcytometry testing currently requires enormous labor.

The present invention efficiently prepares a measurement sample andanalyzes measurement target particles in the sample with high accuracy.

A first aspect of the invention relates to a sample preparationapparatus 1. The sample preparation apparatus 1 includes a measurementunit 2 configured to measure a sample containing particles acquired froma sample container 10 and detect measurement target particles in thesample, a sample preparation unit 3 capable of adjusting theconcentration of measurement target particles in a sample acquired froma sample container 10 and configured to prepare a measurement sample bymixing a sample and any of a plurality of types of particle detectionreagents including a particle labeling substance, and a control unit 4for controlling the sample preparation unit 3 so as to generateconcentration information of the measurement target particles in thesample in the sample container 10 based on the measurement data of themeasurement unit 2 and adjust the concentration of the measurementtarget particles in the sample acquired from the sample container 10according to the generated concentration information and the type ofparticle detection reagent used for preparing the measurement sample.

A second aspect of the invention relates to a sample preparationapparatus 1. The sample preparation apparatus 1 includes a measurementunit 2 configured to measure a sample containing particles acquired froma sample container 10 and detect measurement target particles in thesample, a sample preparation unit 3 configured to prepare a measurementsample by mixing a sample acquired from the sample container 10 and anyof a plurality of types of particle detection reagents including aparticle labeling substance, and a control unit 4 for controlling thesample preparation unit 3 so as to acquire the number of measurementtarget particles in the sample in the sample container 10 based on themeasurement data of the measurement unit 2 and prepare a number ofmeasurement samples based on the acquired number of measurement targetparticles and the type of particle detection reagent.

A third aspect of the invention relates to a sample preparation method.The sample preparation method according to this aspect automaticallyexecutes a step of generating concentration information of a measurementtarget particle in a sample, a step of acquiring type information of aparticle detection reagent used for preparing a measurement sample, astep of automatically adjusting the concentration of the measurementtarget particles in the sample used for preparation of the measurementsample according to the acquired type information and generatedconcentration information, and a step of preparing a measurement sampleby mixing a particle detection reagent with the sample in which theconcentration of measurement target particles has been adjusted.

A fourth aspect of the invention relates to a sample preparation method.The sample preparation method according to this aspect automaticallyexecutes a step of measuring the number of measurement target particlescontained in a sample in a sample container, a step of determining thenumber of measurement sample to be prepared according to the type ofparticle detection reagent and the measured number of measurement targetparticles, and a step of preparing a measurement sample by mixing thesample with the particle detection reagent based on the determinednumber.

A fifth aspect of the invention relates to a sample preparation system1′. The sample preparation system 1′ according to this aspect includes ameasurement device 2′ for measuring a sample containing particlesacquired from a sample container 10 and detecting measurement targetparticles in the sample, a sample preparation apparatus 3′ capable ofadjusting the concentration of the measurement target particles in thesample acquired from the sample container 10 which prepares ameasurement sample by mixing the sample with any one of a plurality ofkinds of particle detection reagents including a particle labelingsubstance, and a control device 4′ that is connected to the measurementdevice 2′ and the sample preparation device 3′ and is configured tocontrol the sample preparation apparatus 3′ so as to generateconcentration information of the measurement target particles in thesample in the sample container 10 based on the measurement data of themeasuring device 2′, and adjust the concentration of measurement targetparticles in the sample acquired from the sample container 10 accordingto the type of particle detection reagent used in the preparation of themeasurement sample and the generated concentration information.

A sixth aspect of the invention relates to a particle analyzer 100. Theparticle analyzer 100 according to this aspect includes a firstmeasurement unit 2 that measures a sample including particles acquiredfrom the sample container 10 and detects measurement target particles inthe sample, a sample preparation unit 3 capable of adjusting theconcentration of measurement target particles in a sample acquired froma sample container 10 and which prepares a measurement sample by mixingthe sample and any one of a plurality of types of particle detectionreagents including a particle labeling substance, a control deviceconfigured to control the sample preparation unit 3 so as to generateconcentration information of the measurement target particles in thesample in the sample container 10 based on the measurement data of thefirst measurement unit 2 and adjust the concentration of the measurementtarget particles in the sample acquired from the sample container 10according to the generated concentration information and the type ofparticle detection reagent to be used in the preparation of themeasurement sample, a second measurement unit 2 configured to measurethe measurement sample prepared by the sample preparation unit 3 anddetect the measurement target particles in the measurement sample, andan analysis unit 5 for analyzing the measurement target particles basedon the measurement data of the second measurement unit 2. Note that thefirst measurement unit and the second measurement unit may beintegratedly configured by the same device or may be configuredseparately with different devices.

According to the first to sixth aspects of the invention, after ameasurement is performed to detect measurement target particles in asample and concentration information of the measurement target particlesin the sample is generated based on the measurement data, theconcentration of the measurement target particles in the sample to besupplied to the sample preparing unit is adjusted according to theconcentration information and the particle detecting reagent used forpreparation of the measurement sample, and the specimen preparation unitthen prepares the measurement sample. Therefore, it is possible toefficiently prepare a measurement sample including the measurementtarget particles at a concentration suitable for the particle detectionreagent, and it is possible to analyze the measurement target particlein the sample with high accuracy.

According to the invention, it is possible to improve the efficiency ofpreparing a measurement sample performed based on the particle detectionreagent and the concentration information of the measurement targetparticles contained in the sample, and to accurately analyze themeasurement target particles in the sample.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a block diagram of a sample preparation apparatus;

FIG. 2 is a brief diagram showing an optical system of a flow cytometer;

FIG. 3 is a schematic configuration diagram of a reagent preparationdevice;

FIG. 4 is a cross-sectional view of a sample preparation unit;

FIGS. 5A and 5B are partially enlarged views of a sample preparationunit;

FIG. 6 is a plan view of a centrifugal separation unit;

FIG. 7 is a partial cross-sectional view of a sample preparation unit;

FIG. 8 is a cross-sectional view of a modification example of a samplepreparation unit;

FIG. 9 is a block diagram of an analysis unit;

FIG. 10 is a flowchart showing a summary of an operation procedure ofprocessing by a control unit;

FIG. 11 is a flowchart showing an operation procedure of processing by acontrol unit;

FIG. 12A is a flowchart showing the operation procedure of the processof a first example of S19 of FIG. 11.

FIG. 12B is a flowchart showing the operation procedure of the processof a second example of S19 of FIG. 11.

FIG. 13A is an example of information concerning preparation of ameasurement sample when preparing a measurement sample by dispensing allof a plurality of cell detection reagents into one dispensing tube;

FIG. 13B is an example of information relating to preparation of ameasurement sample when preparing a measurement sample by dispensing aplurality of cell detection reagents into different dispensing tubes,respectively;

FIG. 14 is a diagram showing an example of measurement data obtained bymeasuring a sample in a sample container and detecting measurementtarget particles in the sample;

FIG. 15 is a diagram showing an example of a method for preparingmeasurement samples for samples A to C; and

FIG. 16 is a block diagram of a sample preparation system.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Hereinafter, embodiments of the invention will be described in detailwith reference to the accompanying drawings. The sample preparationapparatus prepares a measurement sample suitable for analysis of asample that includes particles. The particle analyzer also prepares ameasurement sample suitable for analysis of a sample that includesparticles, and optically analyzes the prepared measurement sample,whereby the number or type of particles contained in the sample isidentified, or both. Specifically, for example, a sample preparationapparatus and a particle analyzer are used to detect hematopoietic stemcells contained in umbilical cord blood or bone marrow. The samplepreparation apparatus and the particle analyzer are used for determiningwhether a sample contains abnormal cells such as hematopoietic tumorcells such as leukemia cells, and cancer cells such as lung cancercells.

The measurement target particles may be, for example, artificialparticles such as metal particles or plastic particles. The particlealso may be a biological component other than cells such as a cylinder,and may be a cell such as a microorganism, an animal cell, a plant cellor the like. The sample containing particles is not limited insofar asit is a liquid containing particles (which may be a stock solution or adilute solution). Preferably, the particle is, for example, a cellculture sample or a biological sample. A cell culture sample is, forexample, a sample containing cells cultured in vitro. Examples ofbiological samples are peripheral blood, umbilical cord blood, bonemarrow, cerebrospinal fluid, ascites, pleural effusion, interstitialfluid, urine and the like. Preferable examples of biological samples areperipheral blood, umbilical cord blood or bone marrow.

The particle detection reagent contains at least one particle labelingsubstance per reagent. The particle labeling substance is not limitedinsofar as particles can be detected. The particle detection reagent ispreferably a cell detection reagent. The particle labeling substancealso is preferably a cell labeling substance. The particle detectionreagent preferably contains at least one cell labeling substanceselected from a group including one or more nucleic acid labelingsubstances that label nucleic acids and one or more protein labelingsubstances that label proteins. Preferred examples of the nucleic acidlabeling substance are ethidium bromide (EB), acridine orange (AO),propidium iodide (PI), 7-amino-actinomycin D (7-AAD),4′,6-diamidino-indole (DAPI), Hoechst33342(2′-(4-ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5′-bi-1H-benzimidazoletrihydrochloride), ethidium homodimer-1, ethidium homodimer 2, ethidiummonoazide, trimethylenebis[[3-[[4-[[(3-methylbenzothiazol-3-ium)-2-yl]methylene]-1,4-dihydroquinoline]-1-yl]propyl]dimethylaminium].tetraiodide(TOTO-1), 4-[(3-(TO-PRO-1), N,N,N′,N′-tetramethyl-N, N′-bis[3-[4-[3-[(3-methylbenzothiazol-3-ium)-2-yl]-2-propenylidene]-1,4-dihydroquinolin-]-1,3-propanediaminium tetraiodide (TOTO-3) or2-[3-[[1-[3-(trimethylamino)propyl]-1,4-dihydroquinolin]-4-ylidene]-1-propenyl]-3-methylbenzothiazol-3-ium-diiodide(TO-PRO-3), or fluorescent dyes represented by the following structuralformula (IV).

In the formula, R₁ and R₄ are alkyl groups having a hydrogen atom, analkyl group, a hydroxy group, an alkyl group having an ether group, analkyl group having an ester group, or a benzyl group which may have asubstituent; R2 and R3 are hydrogen atoms, a hydroxyl group, a halogen,an alkyl group, an alkenyl group, an alkynyl group, or an alkoxy group;Z is a sulfur atom, an oxygen atom, or a carbon atom having a methylgroup; n is 0 or 3; X⁻ is an anion.

In the structural formula (IV), one or another of the R₁ and R₄ is analkyl group having 6 to 18 carbon atoms, and the other is preferably ahydrogen atom or an alkyl group having less than 6 carbon atoms. Thealkyl group having 6 to 18 carbon atoms is preferably an alkyl grouphaving 6, 8 or 10 carbon atoms. As the substituent of the benzyl groupof R₁ and R₄, an alkyl group having 1 to 20 carbon atoms, an alkenylgroup having 2 to 20 carbon atoms, or an alkynyl group having 2 to 20carbon atoms can be mentioned, and a methyl group or an ethyl group ispreferable. R₂ and R₃ include an alkenyl group having 2 to 20 carbonatoms. R₂ and R₃ include an alkoxy group having 1 to 20 carbon atoms,and preferably a methoxy group or an ethoxy group in particular. As theanion of X⁻, a halogen ion such as F⁻, Cl⁻, Br⁻, I⁻ or CF 3, CF₃SO₃ ⁻,BF 4⁻ and the like may be considered.

Examples of a nucleic acid labeling substance include a fluorescentlylabeled nucleic acid probe and a fluorescently labeled nucleotide.Examples of a protein labeling substance include a fluorescent substanceis labeled, and a protein capable of binding to a target protein or avitamin capable of binding to an objective protein. Examples of aprotein binding to a target protein include a ligand, an antibody, alectin or a lipid binding protein (preferably a phospholipid-bindingprotein) which binds to a receptor. The fluorescent substance is notlimited insofar as it can be detected by the measurement unit 2. Thefluorescent substance is preferably a substance that can be used forflow cytometry.

The particle detection reagent set in the reagent setting unit 35 of thereagent preparation apparatus 1 to be described later preferablycorresponds to a plurality of particle labeling substances.“Corresponding to a particle labeling substance” may refer to a case inwhich a plurality of particle labeling substances are contained in oneparticle detection reagent, or a case in which a plurality of particledetection reagents contain one or more particle labeling substances.

It is preferable that each particle detection reagent or each particlelabeling substance is mixed with particles contained in the sample at anappropriate mixing ratio. The ratio also may be different for eachparticle detection reagent or each particle labeling substance. Forexample, when a plurality of types of particles are contained in onesample, the mixing ratio is determined in consideration of theproportion of measurement target particles.

In the following embodiments, the measurement target particles aredescribed as cells, but the sample preparation apparatus and theparticle analyzer are not limited only to analysis of cells.

Sample Preparation Apparatus and Particle Analyzer Structures

FIG. 1 shows a schematic configuration of the particle analyzer 100 ofthe embodiment. The particle analyzer 100 of the embodiment includes asample preparation apparatus 1 and an analysis unit 5. The samplepreparation apparatus 1 according to the embodiment includes ameasurement unit 2 that performs preliminary measurement of a sample anda main measurement of a measurement sample, a sample preparation unit 3that adjusts the concentration of a sample and prepares a measurementsample, and a control unit 4 for controlling the measurement unit 2 andthe sample preparation unit 3. The analysis unit 5 analyzes themeasurement data obtained by the main measurement of the measurementunit 2 and the like.

Measurement Unit Structure

The measurement unit 2 functions as a pre-measurement unit thatpre-measures the sample and detects the number of measurement targetcells contained in the sample. The measurement unit 2 of the embodimentalso functions as a main measurement unit that performs a mainmeasurement of the measurement sample and detects information on thecharacteristics of the measurement target cells for cell analysis by theanalysis unit 5. A flow cytometer is employed as the measurement unit 2of the embodiment.

FIG. 2 is a schematic diagram showing the optical system of the flowcytometer configuring the measurement unit 2. The flow cytometerincludes a flow cell 20 through which a sample passes, light sources 21Aand 21B for irradiating light on the sample passing through the flowcell 20, and a light receiving element 22A to 22F for detecting opticalinformation of the light given off from the particles in the sample andconverting the optical information into electric signals that are outputas detection signals.

The optical information is information included in one or two or morelight wavelength spectra emitted from particles. The light wavelengthspectra includes individual light wavelengths, light wavelength regionsincluded in the light wavelength spectrum, light intensities of therespective light wavelengths, and light intensities in the lightwavelength region.

The light emitted from the light source 21A irradiates the flow cell 20via the collimator lens 23A, the dichroic mirror 24A, and the condenserlens 25A. The forward scattered light of the light derived from theparticles passing through the flow cell 20 is condensed by the condenserlens 25B and enters the light receiving element 22A via the beam stopper26, the pin hole plate 27A, and the band pass filter 28A.

On the other hand, side scattered light and lateral fluorescent lightgiven off from the particles passing through the flow cell 20 arecollected by the condenser lens 25C. The side scattered light enters thelight receiving element 22B through the dichroic mirrors 24B to 24D, thepin hole plate 27B and the band pass filter 28B. The side fluorescentlight having a wavelength of 520 nm or more and 542 nm or less passesthrough the dichroic mirrors 24B and 24C, then is reflected by thedichroic mirror 24 D, and enters the light receiving element 22C via thepinhole plate 27C and the bandpass filter 28C. The side fluorescentlight having a wavelength of 570 nm or more and 620 nm or less passesthrough the dichroic mirror 24B and is reflected by the dichroic mirror24C, and passes through the pinhole plate 27D and bandpass filter 28Dand impinges on the light receiving element 22D. Side fluorescent lighthaving a wavelength of 670 nm or more and 800 nm or less is reflected bythe dichroic mirror 24B, passes through the dichroic mirror 24E, andenters the light receiving element 22E via the pinhole plate 27E and thebandpass filter 28E.

The light emitted from the light source 21B is irradiated on the flowcell 20 via the collimating lens 23B, the dichroic mirror 24A, and thecondensing lens 25A. The side fluorescent light of the light given offfrom the particles passing through the flow cell 20 is collected by thecondenser lens 25C. The side fluorescent light of 662.5 nm or more and687.5 nm or less is reflected by the dichroic mirror 24B, reflected bythe dichroic mirror 24E, and then enters the light receiving element 22Fvia the pinhole plate 27F and the bandpass filter 28F.

For example, a laser diode with a wavelength of 488 nm is used for thelight source 21A, and a laser diode with a wavelength of 642 nm is usedfor the light source 21B. A sheath flow cell is used for the flow cell20. A photodiode is used for the light receiving element 22A thatreceives the forward scattered light, an avalanche photodiode (APD) isused for the light receiving element 22B that receives the sidescattered light, and a photomultiplier tube (Photo Multiplier Tube, PMT)is used for the light receiving elements 22C to 22F that receive theside fluorescent light. In FIG. 2, note that although the flow cytometerhas six light receiving elements 22A to 22F and the four light receivingelements 22C to 22F detect optical information of four lights havingdifferent peak wavelengths derived from a dye bonded to particles in asample, the invention is not limited to this arrangement inasmuch as itis also possible to provide three or more light receiving elementswherein at least two or more of the three or more light receivingelements detect optical information of light derived from at least twodyes having different peak wavelengths.

The number of light sources may be one, or two or more. For example, thenumber of light sources can be selected between integers from 1 to 10.The light source is selected according to the wavelength region of lightderived from the dye bound to the particle. When the light sources are 2or more, it is preferable that these light sources emit light havingdifferent peak wavelengths. two or more light sources are preferablebecause it is possible to separate and detect fluorescence with highaccuracy as compared to when there is only one light source. On theother hand, when two light sources are used, it is possible to separateand detect a plurality of fluorescence by shifting the light emissiontiming from each light source. By using a dye suitable for the peakwavelength of light from each light source, it is also possible toreduce the overlapping portion of the respective wavelength regions ofthe plurality of fluorescences by shifting the light emission timingfrom each light source. The light source is not limited insofar as lightin the wavelength region for detection is emitted from the particle. Forexample, one or more light sources selected from a group including ahalogen lamp, an LED (Light Emitting Diode) lamp, a gas laser, and alaser diode (semiconductor laser) can be used as a light source. Thenumber of photodiodes, dichroic mirrors, and bandpass filters can bevaried according to the number of peak wavelengths of light originatingfrom the particles. The types of the photodiode, the dichroic mirror,and the bandpass filter can also be selected according to the peakwavelength of light derived from the particle, the wavelength region,and the intensity thereof.

Detection signals output from the light receiving elements 22A to 22Fare amplified by a preamplifier (not shown) and sent to the signalprocessing unit 6 (shown in FIG. 1). The signal processing unit 6includes a signal processing circuit that performs signal processingnecessary for the detection signal output from the measurement unit 2.

Sample Preparation Unit Structure

As shown in FIGS. 1 and 3 to 7, the sample preparation unit 3 adjuststhe concentration of the measurement target cells in the sample used toprepare the measurement sample, the prepares a measurement sample fromthe sample and one or more cell detection reagents selected a pluralityof cell detection reagents. Note that the configuration of the samplepreparation unit 3 shown below is merely an example, and the samplepreparation unit 3 is not limited to the following configuration.

The sample preparation unit 3 includes a sample dispensing unit 30A, areagent dispensing unit 30B, a sample container setting unit 31, adispensing tube setting unit 32, a dispensing tube storing unit 33, apipette tip storing unit 34, a reagent setting unit 35, a tube transferunit 36, a liquid amount detection unit 37, a centrifugal separationunit 38, and a receiving unit 39. Note that in FIGS. 3 to 6 the XYZ axesare orthogonal to one another, and the X axis indicates the horizontaldirection, the Y axis indicates the front and rear direction, and the Zaxis indicates the vertical direction.

Each of the sample dispensing units 30A and the reagent dispensing unit30B has a nozzle 300. For example, a disposable plastic pipette tip 301is mounted on the nozzle 300. Note that it is not always necessary touse the pipette tip 301. The sample dispensing unit 30A suctions anddischarges a predetermined amount of sample from the sample container 10of the sample container setting unit 31 by suctioning and ejecting thesample through the pipette tip 301 of the nozzle 300, and discharges thesample to the dispensing tube 11. The nozzle 300 suctions and dischargesthe liquid through the pipette tip 301, so that the reagent dispensingunit 30B suctions a predetermined amount of the reagent from the reagentcontainer 350 of the reagent setting unit 35 and supplies the reagent tothe dispensing tube 11. The amount (volume) of the sample or reagentsuctioned by the nozzle 300 can be acquired from a flow rate sensor (notshown) provided in each dispensing unit 30A, 30B. Each of the dispensingunits 30A and 30B includes a nozzle transfer unit 302 that moves thenozzle 300 in the X, Y, and Z axis directions. The nozzle transfer unit302 moves the nozzle 300 by driving a motor (not shown).

In the sample dispensing unit 30 of the embodiment, the nozzle 300 isprovided with a liquid surface detection sensor (not shown) fordetecting the liquid level of the sample in the sample container 10 whenit is positioned directly above the sample container 10 as a liquidamount detection unit 37 for detecting the amount of the samplecontained in the sample container 10. By detecting the liquid level ofthe sample in the sample container 10, it is possible to calculate theliquid amount (sample amount) of the sample contained in the samplecontainer 10.

A sample container 10 containing a sample collected from a subject isset in the sample container setting unit 31. A plurality of samplecontainers 10 can be set in the sample container setting unit 31. In thedispensing tube setting unit 32, a dispensing tube 11 for accommodatinga sample and a prepared sample used for measurement by the measurementunit 2 is set in the sample container setting unit 31. A plurality ofdispensing tubes 11 can be set in the dispensing tube setting unit 32.

The sample container setting unit 31 includes a sample temperatureadjusting unit 310 such as a warming device or a cooling device. Thetemperature of the sample accommodated in the sample container 10 can beset to a temperature suitable for the sample by the sample temperatureadjusting unit 310. The cooling device is configured by, for example, aPeltier element, a compressor, or other cooling device. The heatingdevice is configured by, for example, a block heater or other heater.The temperature inside the sample container setting unit 31 is measuredby a temperature sensor (not shown).

A plurality of dispensing tubes 11 are stocked in the dispensing tubestorage unit 33. The dispensing tube 11 of the dispensing tube storageunit 33 is transported to the dispensing tube setting unit 32 by thetube transfer unit 36 and set.

A plurality of pipette tips 301 to be attached to the nozzle 300 arestocked in the pipette tip storage unit 34. Pipette tips 301 ofdifferent sizes are stocked in the pipette tip storage section 34. Apipette tip 301 of either a large or small size can be attached to thenozzle 300, and a pipette tip 301 of a size corresponding to thedispensing amount of a sample or a reagent is attached to the nozzle300. Note that a used pipette tip 301 is discarded in the discard unit340.

The reagent setting unit 35 is a reagent storage having a plurality ofreagents, and a plurality of reagent containers 350 containing reagentsare set inside the reagent setting unit 35. A plurality of openings 351are formed in the reagent setting unit 35, respectively, and a reagentcontainer 350 is set below the opening 351. The reagent can be suctionedby allowing the nozzle 300 of the reagent dispensing unit 30B to enterthe reagent container 350 from the opening 351.

A plurality of cell detection reagents are set in the reagent settingunit 35. It is preferable that at least one cell labeling substancecontained in each cell detection reagent is different among the celldetection reagents. The combination of a plurality of cell detectionreagents set in the reagent setting unit 35 at one time, for example,may be a combination covering a group of cell labeling substancescapable of detecting a group of cell markers necessary for specifyingthe type of leukemia. The cell detection reagent also may contain abuffer solution in addition to the cell labeling substance. The buffersolution also may contain a salt such as sodium chloride. It ispreferable that the salt is added so as to become isotonic within thecell when preparing the measurement sample. The cell detection reagentalso may contain cell fixing components such as methanol,paraformaldehyde and the like. The cell detection reagent also maycontain RNase or the like.

In addition to the cell detection reagent, other reagents necessary forthe preparation of the measurement sample according to the analysis ofthe measurement target cells, such as, for example, a hemolytic agent, awashing solution, a cell membrane penetrating agent, a diluting liquid,a cell fixing component, RNase, also can be set in the reagent settingunit 35. Note that the diluting liquid contains at least a buffersolution and may optionally contain a salt such as sodium chloride. Thesalt may be added so as to be isotonic within the cell when preparingthe measurement sample. The diluting liquid may contain a hemolyticagent that causes hemolysis of erythrocytes. It is preferable that thehemolytic agent does not dissolve nucleated cells such as white bloodcells. The hemolytic agent is preferably an aqueous solution containinga surfactant, citrate buffer, HEPES, phosphate buffer or the like. Thesurfactant may be any of an anionic surfactant, a cationic surfactant, abipolar surfactant, a nonionic surfactant, a natural surfactant, and thelike. One hemolytic agent also may contain a plurality of kinds ofsurfactants. More preferably, a hemolytic agent containing a cationicsurfactant and an organic acid and having a pH in the range of 4.5 to11.0, a hemolytic agent including a nonionic surfactant and an organicacid and having a pH in the range of 4.5 to 11.0, and a hemolytic agentcontaining an anionic surfactant and a natural surfactant and having apH in the range of 4.5 to 11.0 can be mentioned. The pH range ispreferably pH 6.0 to 8.0. These hemolytic agents also may containalcohols such as methanol, ethanol, phenoxyethanol and the like,fixative solutions such as formaldehyde, chelating agents, sodium azideand the like. More specific hemolytic agents include ammonium chloridehemolytic agents (pH 7.3, ammonium chloride 1.68 M, potassium hydrogencarbonate 100 mM, EDTA 2 K 0.82 mM), cell fixing hemolytic agent (pH 7,formaldehyde 3, 10%, methanol 3.5%, diethyleneglycol 30%, citric acid100 mM), cell membrane permeable hemolytic agent (pH 7.3, phenoxyethanol1% or less, saponin 1% or less, N-laurylsarcosine sodium salt 1% orless, sodium azide 1% or less can be mentioned. The cell membranepermeable hemolytic agent also can be used as a cell membranepenetrating agent when detecting intracellular proteins.

Each reagent container 350 set in the reagent setting unit 35 isprovided with a barcode, tag, or the like storing information on eachreagent. The information on the reagent includes identificationinformation (ID) for specifying the reagent. The information on thereagent also may include the name of the measurement item or the like.The reagent setting unit 35 includes a receiving unit 39 (shown inFIG. 1) such as an RFID reader, barcode reader capable of readingbarcodes and the like, such that the information relating to the celldetection reagents and reagents other than the cell detection reagentset in the reagent setting unit 35 can be acquired and by reading abarcode or the like attached to the reagent container 350.

Note that in order to keep the temperature of the reagent in eachreagent container 350 at a desired temperature, the reagent setting unit35 may be provided with an internal reagent temperature adjusting unit(not shown). The reagent temperature adjusting unit is a cooling devicecapable of cooling the reagent in the reagent container 350 and/or aheating device capable of heating the reagent in the reagent container350. The temperature inside the reagent setting unit 35 can be measuredby a temperature sensor (not shown).

The tube transfer unit 36 includes a gripper 360 that holds thedispensing tube 11, and a moving unit 361 that moves the gripper 360 inthe X, Y, and Z axis directions. The moving unit 361 moves the gripper360 by driving a motor (not shown). The dispensing tube 11 of thedispensing tube setting unit 32 is transported to the centrifugalseparation unit 38 and the measurement unit 2 by being moved while beingheld by the gripping 360.

The centrifugal separation unit 38 performs a preparation process forpreparing a measurement sample from the sample and a predeterminedreagent. The centrifugal separator 38 also performs a concentrationadjustment process to adjust the concentration of measurement targetcells in the sample by centrifugal separation. That is, in theembodiment, the centrifugal separation unit 38 functions as aconcentration adjustment unit.

The centrifugal separation unit 38 is provided in a housing sectionprovided with a cylindrical bottom tank body 390, a lid body 391covering the upper opening of the tank body 390, and a temperatureadjustment tank 392 provided at the lower part of the tank body 390. Thelid 391 is attached to the tank body 390 via a hinge 393. The lid body391 is formed with a first entry port 394 for entering into the tankbody 390 from above. The first entry port 394 allows the nozzle 300 ofeach dispensing section 30A and 30B to enter into the tank body 390, andthe nozzle 300 enters the tank body 390 through the first entry port 394to discharge the sample or reagent into the dispensing tube 11 held inthe centrifugal separation unit 38 or to suction the liquid in thedispensing tube 11.

The tank body 390 is provided with opening/closing device 395 foropening and closing the first entry port 394. The opening/closing device395 of the embodiment is composed of a shutter 395 a for closing thefirst entry port 394, a drive unit 395 b for opening and closing theshutter 395 a, and a connection unit 395 c for connecting the shutter395 a and the drive unit 395 b. The driving unit 395 b is, for example,a solenoid, and moves the shutter 395 a in the Y-axis direction. Asshown in FIG. 5 (a), when the first entry port 394 is closed by theshutter 395 a, the inside of the tank body 390 is substantially sealed,and temperature change in the tank body 390 can be suppressed. As shownin FIG. 5 (b), when the shutter 395 a moves and the first entry port 394is opened, the nozzle 300 also can be advanced into the tank body 390from the first entry port 394.

Note that, as shown in FIG. 4, the tank body 390 has a second entry port396 for entering into the tank body 390 from behind. The second entryport 396 allows the gripper 360 of the tube transfer unit 36 to enterinto the tank body 390, so that the gripper 360 enters the tank body 390through the second entry port 396 to set the dispensing tube 11 in theholding part 380 of the centrifugal separation unit 38, and to removethe dispensing tube 11 from the holding part 380 of the centrifugalseparation unit 38. The shutter 395 a is formed in an L shape in crosssection so that the second entry port 396 can be closed together withthe first entry port 394.

The centrifugal separation unit 38 includes a plurality of holding parts380 for holding the dispensing tube 11, and a rotor 381 on which aplurality of holding parts 380 are attached on the outer circumference.Centrifugal separation is performed on the liquid in the dispensing tube11 held by each holding part 380 by the rotation of the rotor 381. Therotor 381 rotates around a rotation shaft 382 rotatably supported at thebottom of the tank body 390 as the center of rotation. A first pulley383 is provided under the rotation shaft 382. A second pulley 386 isprovided on a rotation shaft 385 of a motor 384 that rotationally drivesthe rotation shaft 382, and an endless belt 387 is wound between thefirst pulley 383 and the second pulley 386. The rotation of the motor384 is transmitted to the rotation shaft 382 through the first pulley383, the endless belt 387, and the second pulley 386 to rotate the rotor381. The motor 384 is disposed outside the sample preparation unit 8.

A temperature adjustment tank 392 is partitioned by the tank body 390and the bottom portion of the tank body 390. Through holes 397 a and 397b are formed in the bottom of the tank body 390, and the tank body 390and the temperature adjustment tank 392 communicate with each otherthrough the through holes 397 a and 397 b.

A temperature adjustment device 398 for adjusting the atmospherictemperature inside the tank body 390 is provided in the temperatureadjustment tank 392. The temperature adjusting device 398 has, forexample, a Peltier element 398 a. In the temperature adjustment tank392, convection generating units 399 a and 399 b are provided togenerate convection circulating through the tank body 390 and thetemperature adjustment tank 392 via the through-holes 397 a and 397 b.The convection generating units 399 a and 399 b are configured by, forexample, a fan or the like. The interior of the tank body 390 can beheated to maintain a temperature higher than room temperature, theinterior of the tank body can be cooled to maintain a temperature lowerthan room temperature, and the interior of the tank body can bemaintained at a constant temperature approximately equal to roomtemperature by moving cold or heat from the temperature adjustment tank392 to the tank body 390 by the air flow generated by the convectiongenerating units 399 a and 399 b. In the tank body 390, a temperaturesensor 400 is provided to monitor the temperature inside the tank body390. Note that the temperature sensor 400 also may be provided in thetemperature adjustment tank 392.

In the embodiment, the convection generating units 399 a and 399 bpositioned near the convection generating unit 399 a moves the cold orheat emitted from the temperature adjustment device 398 to the throughhole 397 b located away from the temperature adjustment device 398 bymoving the convection flowing from the through hole 397 a located abovethe convection generating unit 399 a in a horizontal direction insidethe temperature adjustment tank 392. The convection generating unit 399b is located below the through hole 397 b and changes the horizontal airflow generated by the convection generating unit 399 a into an upwardair flow directed toward the upper through hole 397 b. In this way thecold or heat emitted from the temperature adjustment device 398 can beefficiently fed from the through hole 397 b into the tank body 390.

In the embodiment, a duct 401 is provided behind the temperatureadjustment tank 392. The duct 401 is for discharging cold exhaust heator warm exhaust heat generated by the temperature adjustment device 398to the outside of the sample preparation unit 8. The duct 401 includes afan 402 for letting cold exhaust heat or warm exhaust heat to the rearand an exhaust port 403 for discharging the air current generated by thefan 402 to the outside of the duct 401. The duct 401 is arranged belowan external processing unit 37.

Note that the temperature adjustment tank 392 is not necessarilyprovided, as shown in FIG. 8. In this case, the temperature adjustingdevice 398 is provided inside the tank body 390, preferably on the rearside in the tank body 390, and the duct 401 is provided behind the tankbody 390. The convection generating unit 399 a is arranged in thevicinity of the temperature adjusting device 398 so as to generate ahorizontal air current in the tank body 390.

The sample preparation unit 3 may be provided with an externalprocessing unit 60 in addition to each unit described above. In theembodiment, the external processing unit 60 performs processing at roomtemperature without temperature adjustment, and is provided with adisposal unit 600 for discarding the supernatant in the dispensing tube11, and a stirring unit 601 for stirring the liquid in the tube 11. Thedisposal unit 600 discards the supernatant in the dispensing tube 11 bydecanting by tilting the dispensing tube 11 held by the gripper 360 viaa moving device 361, or suctioning the supernatant through the nozzle300 of the dispensing units 30A and 30B. The stirring unit 601 isconfigured by, for example, a vortex mixer that stirs and shakes theliquid in the dispensing tube 11 held by the gripper 360. Note that theexternal processing unit 60 may be configured to perform processingother than the disposal of the supernatant in the dispensing tube 11 andthe stirring of the liquid in the dispensing tube 11 at roomtemperature.

Control Unit Structure

As shown in FIG. 1, the control unit 4 includes a processor 40, a memory41 used for reading a control program recorded in the storage unit 42and used for a work area for data processing of the processor 40, and astorage unit 42 for recording various control programs and various datafor controlling the operation of each unit such as the measurement unit2 and the sample preparation unit 3. The memory 41 is configured by aRAM (Random Access Memory). The storage unit 42 is configured by a ROM(read only memory), a hard disk, or the like.

For example, the control unit 4 acquires detection signals output fromthe light receiving elements 22A to 22F of the measurement unit 2 viathe signal processing unit 6, and records them in the storage unit 42.

The control unit 4 also controls the operations of the sample dispensingunit 30A of the sample preparation unit 3, the nozzle 300 of the reagentdispensing unit 30B, and the nozzle transfer unit 302 to perform themovement of the nozzle 300 and the suction and discharge the liquidsample and reagent and the like.

The control unit 4 also controls the operation of the temperatureadjustment unit 310 of the sample container setting unit 31 of thesample preparation unit 3 and the operation of the temperatureadjustment unit (not shown) of the reagent setting unit 35 based ondetection signals of a temperature sensor (not shown), and adjusts thetemperatures of the sample container setting unit 31 and the reagentsetting unit 35.

The control unit 4 also controls the operation of the tube transfer unit36 of the sample preparation unit 3 to transfer and hold the dispensingtube 11.

The control unit 4 also controls the opening/closing unit 395 of thesample preparation unit 3 to open and close the entry ports 394 and 396.

The control unit 4 also controls the centrifugal separation unit 38 ofthe sample preparation unit 3 and performs centrifugal separationprocessing.

The control unit 4 also controls the temperature adjustment unit 398 andthe convection generation units 399 a and 399 b of the samplepreparation unit 3 based on the detection signal of the temperaturesensor 400 to adjust the temperature of the centrifugal separation unit38.

The control unit 4 also controls the operation of the fan 402 of thesample preparation unit 3 to exhaust the cold/warm heat of thecentrifugal separation unit 38.

The control unit 4 also acquires a detection signal from the liquidlevel detection sensor as the liquid amount detection unit 37 of thesample preparation unit 3, and records the data in the storage unit 42.

The control unit 4 also acquires identification information (ID) of eachreagent (a cell detection reagent and a reagent other than the celldetection reagent) read by the receiving unit 39 of the reagent settingunit 35 of the sample preparation unit 3. Then, on the basis of the readidentification information (ID) of each reagent, data indicating whichreagent is set at which position of the reagent setting unit 35 isrecorded in the storage unit 42. Further, information can be readrelated to the preparation of a measurement sample corresponding to eachtest item to be tested using each cell detection reagent correspondingto the identification information (ID) recorded in the storage unit 42based on the identification information (ID) of each cell detectionreagent that has been read, and the sample preparation unit 3 can becontrolled based on the information related to the preparation of themeasurement sample to perform the concentration adjustment process ofthe measurement target particle in the sample and the preparationprocess of the measurement sample. Note that information related to thepreparation of the measurement sample is determined independently by theuser according to the measurement sample and recorded together with thetype of the cell detection reagent and the like in a file referred to asa work list.

The manner of preparing the measurement sample differs depending onmeasurement items (for example, analysis of DNA amount, kind of antigenand the like), since it is necessary to prepare a measurement samplesuitable for the cell analysis for each measurement item. Theinformation on the preparation of the measurement sample includesinformation on the preparation of the measurement sample according tothe measurement item of the measurement target cells, and a measurementsample suitable for cell analysis can be prepared by preparing themeasurement sample based on the information on preparation of themeasurement sample.

Information related to the preparation of the measurement sample alsomay include information on measurable measurement items and informationon the test items, characteristics of the cell detection reagent usedfor cell analysis (including information on measurement items,information on particle labeling substances to label antibodies and thelike, cross reactivity of the antibodies, measurement items which may bemeasured simultaneously and the like), the type of sample, the number ofcells (necessary number of cells) required for cell analysis using eachcell detection reagent, the ratio of the measurement target cells persample amount, the amount of sample, the type of other reagentsnecessary for preparing the measurement sample, the dispensing amount ofeach reagent, the order of reagents to be dispensed, the temperature atreagent dispensing, the number of measurement samples (pipetting tubes11) required for the measurement and the like.

The control unit 4 also is connected to the analysis unit 5 via acommunication interface 7, and transmits and receives measurement datameasured by the measurement unit 2 and data necessary for processing ofeach unit to and from the analysis unit 5.

Analysis Unit Structure

As shown in FIG. 9, the analysis unit 5 includes a processing unit 50,an input unit 51, and a display unit 52. The processing unit 50 includesa processor 54, a memory 55, a storage unit 56, an input/outputinterface 57, an image output interface 58, and a communicationinterface 59. The processing unit 50 can be configured by ageneral-purpose computer. The memory 55 is configured with a RAM (RandomAccess Memory). The memory 55 is used for reading out the computerprogram recorded in the storage unit 56. The memory 55 also is used as awork area for various data processing of the processor 54. The storageunit 56 is configured by a ROM (read only memory), a hard disk, and thelike. The storage unit 56 records the computer program and variousprocessing data used by the computer program. An operation program forsending operation commands to the control unit 4, receiving andanalyzing the measurement data performed by the measurement unit 2,displaying processed analysis results and the like is installed in thestorage unit 56 (for example, a hard disk).

The input unit 51 is configured by, for example, a touch panel, akeyboard, a mouse, a pen tablet, and the like. The display unit 52 isconfigured by, for example, a display or the like.

Control Unit Operations

As shown in FIG. 10, the control unit 4 performs processes including apre-measurement step S1 for measuring a sample acquired from the samplecontainer 10 and detecting measurement target cells in the sample, ageneration step S2 of generating concentration information of themeasurement target cells in the sample in the sample the samplecontainer 10 based on the measurement data of the pre-measurement step,a determination step S3 of acquiring the information of the type ofparticle detection reagent used for preparing the measurement sample anddetermining the inspection item, an adjustment step S4 of automaticallyadjusting the concentration of measurement target cells in the sample inaccordance with the type of the cell detection reagent used forpreparation of the measurement sample (information related to thepreparation of the measurement sample), a preparation step S5 forpreparing a measurement sample from the sample and the cell detectionreagent, and measurement step S6 for performing the main measurement ofmeasurement sample. Details will be described below.

First Embodiment

A first embodiment of the operation of the control unit 4 will bedescribed with reference to FIG. 11 and FIG. 12A. Note that thefollowing flow is merely an example, and the embodiment is not limitedto the following flow. The first embodiment is an example in which allof a plurality of cell detection reagents necessary for preparing ameasurement sample for measuring each measurement item of apredetermined test item are mixed with a sample in one dispensing tube11 to prepare the measurement sample.

First, in S10 of FIG. 11, the control unit 4 receives input ofidentification information (ID) of the cell detection reagent used forpreparation of the measurement sample. Prior to the measurement, aplurality of cell detection reagents and reagents other than the celldetection reagent necessary for preparation of the measurement sampleare set in the reagent setting unit 35 of the sample preparation unit 3.At this time, the receiving unit 39 of the reagent setting unit 35acquires identification information (ID) of the reagent in the reagentcontainer 350 from the barcode or the like attached to the reagentcontainer 350. Upon receiving the identification information (ID) ofeach reagent, the control unit 4 determines the type of the particledetection reagent used for preparation of the measurement sample basedon the identification information (ID), and proceeds to step S11 to readfrom the storage unit 42 the information relating to the preparation ofthe measurement sample of each measurement item corresponding to thetest item to be tested using the cell detection reagent of thedetermined type. In S12, the control unit 4 also transmits theinformation relating to preparation of the read measurement sample tothe analysis unit 5. Data indicating which reagent is set at whichposition in the reagent setting unit 35 also is recorded in the storageunit 42.

For example, FIG. 13A is an example of information related topreparation of a measurement sample recorded in the storage unit 42 inthe example where the sample is peripheral blood and the examinationitem is regulatory T cell. CD25, CD3 and CD4 shown in FIG. 13A are cellsurface markers, and cell detection reagents 2 and 3 contain antibodiesbinding to these cell surface markers, respectively. The antibody alsois labeled with the fluorescent substance shown in FIG. 13A. FoxP3 is anuclear protein, and cell detection reagent 4 contains a fluorescentlylabeled anti-FoxP3 antibody to the protein. In addition, the informationconcerning the preparation of the measurement sample in FIG. 13Aincludes information on the number of necessary cells, leukocytesnecessary for performing cell analysis using each cell detection reagent1 to 4, the amount of specimen, the type of other reagents required toprepare a measurement sample, the mixing amount of each reagent, theorder of the reagents to be dispensed and the like.

Next, in S13, the control unit 4 controls the sample dispensing unit 30Ato suction a part of the sample in the sample container 10 set in thesample container setting unit 31, by the nozzle 300, and discharge thesuctioned sample to the dispensing tube 11. The sample may be dischargedto the dispensing tube 11 set in the dispensing tube setting unit 32 ormay be discharged into the dispensing tube 11 set in the centrifugalseparation unit 38 by the tube transfer unit 36. Note that, ifnecessary, a predetermined amount of diluting liquid set in the reagentsetting unit 35 is suctioned by the nozzle 300 of the reagent dispensingunit 30B, and discharged to the dispensing tube 11 into which the samplehas been dispensed to be mixed therewith. The control unit 4 alsocalculates the liquid amount of the sample in the sample container 10based on a detection signal from a liquid level detection sensor (notshown) employed as the liquid amount detection unit 37.

Next, in step S14, the control unit 4 controls the tube transfer unit 36to transfer the dispensing tube 11 into which the sample has beendispensed to the measurement unit 2. When the dispensing tube 11 istransported to a predetermined position 29 (shown in FIG. 3) of themeasurement unit 2, the sample is suctioned from the dispensing tube 11by the suction unit 29 a that can move up and down in the verticaldirection (Z direction), and is supplied to the flow cell 20 of themeasurement unit 2. In this way the control unit 4 measures the sampleby flow cytometry analysis using the measurement unit 2 in S15, andcounts the number of measurement target cells in the sample.

Next, in S16, the control unit 4 generates concentration information ofthe measurement target cells in the sample in the sample container 10based on the measurement data of the measurement unit 2. Note that theconcentration of measurement target cells in a sample refers to thenumber of cells to be measured contained in the sample per unit volume.Specifically, the amount of sample suctioned from the sample container10 by the nozzle 300 of the sample dispensing unit 30A for samplemeasurement can be obtained from a flow rate sensor (not shown) providedin the sample dispensing unit 30A, and the concentration information ofthe measurement target cells in the sample can be generated by dividingthe cell number (measurement data) of the measurement target cellscounted by the measurement unit 2 by the suctioned sample volume.

Note that when the measurement unit 2 is a flow cytometer, the number ofmeasurement target cells can be counted based on the detection signal ofthe forward scattered light. When the measurement unit 2 is a flowcytometer, the number of cells of the measurement target cell can becounted in consideration of the proportion of the measurement targetcells in a plurality of kinds of cells included in the sample.Specifically, the ratio of the measurement target cells counted by themeasurement unit 2 is added to the total number of cells counted by themeasurement unit 2, and the number of measurement target cells and theconcentration of the measurement target cells are calculated.

Next, in S17, the control unit 4 determines the test item. Thedetermination of the test item is determined by specifying the test itemincluding the identification information (ID) of each cell detectionreagent by the identification information (ID) of the plurality of celldetection reagents acquired by the receiving unit 39 described above.For example, in the example shown in FIG. 13A, the test item A(regulatory T cell) is determined based on the identificationinformation (ID) 1 to 4 of the cell detection reagents obtained by thereceiving unit 39. In S18, the control unit 4 also transmits thedetermined test item to the analysis unit 5.

Note that the control unit 4 compares the concentration information ofthe measurement target cells in the sample generated in S16 with areference value stored in the storage unit 42, and when theconcentration of the measurement target cell is higher than thereference value, test items corresponding to another cell detectionreagent group set in the reagent setting unit 35 also may be measured.

For example, when the sample is peripheral blood, the control unit 4compares the concentration information of the measurement target cells(preferably nucleated cells) obtained by the preliminary measurementwith the reference value of the number of the cells recorded in thestorage unit 42. The reference value is about 15,000 cells/μl, forexample, when the ample is peripheral blood. A hematopoietic tumor issuspected if the concentration of nucleated cells obtained bypre-measurement exceeds this value. Therefore, if it is determined thatthe concentration of the nucleated cells obtained by the preliminarymeasurement exceeds this value, the control unit 4 further controls thesample preparation unit 3 to prepare a measurement sample for adding ananalysis item to identify hematopoietic tumor. The analysis item foridentifying the hematopoietic tumor may be protein or nucleic acid.

Next, in S19, the control unit 4 determines whether measurement of allmeasurement items is possible. In step S19, the measurement target cellsin the sample in the sample container 10 (the total number of cells) iscalculated based on the concentration information of the measurementtarget cells in the sample generated in S16 and the sample amount in thesample container 10 calculated in S19 of FIG. 12. Next, in step S191,the control unit 4 determines whether the calculated number ofmeasurement target cells in the sample is equal to or larger than thenumber of cells sufficient for measurement of all the measurement itemsof the test item.

For example, in the example of FIG. 13A, the white blood cell countnecessary for measurement of regulatory T cells is, for example, 1×10⁴.Here, as shown in FIG. 14, the leukocyte concentration of sample A is1×10³/Ml from result of the pre-measurement of the sample, and when thesample volume is 1 ml, the number of measurement target cell leukocytesin the sample A is 1×10⁶, therefore, sample A satisfies the measurementconditions of all the measurement items. Since the number of measurementtarget cell leukocytes in sample B is 1×10⁷ cells and 1×10⁵ cells,samples B and C satisfy the measurement conditions of all themeasurement items. Therefore, in this case, S191 in FIG. 12A becomes“YES” and proceeds to S192, the control unit 4 determines allmeasurement items as measurement items. The determination result also isrecorded in the storage unit 42 and transmitted to the analysis unit 5,and the process proceeds to S20 of FIG. 11.

On the other hand, although not shown in the figure, the measurementcondition of the measurement item using the cell detection reagent 4will not be satisfied when the number of leukocytes which aremeasurement target cells in the sample is less than 1×10⁴. In this case,“NO” is obtained in S191 of FIG. 12A and the flow advances to S193, anda notice that the number of cells necessary for measuring themeasurement item cannot be ensured and a measurement item can not bemeasured is sent to the analysis unit 5.

In the case where the number of measurement target cells in the sampleis equal to or larger than the number of cells sufficient formeasurement of all the measurement items, the number of measurementtarget cells in the sample also can be rendered equal to or greater thanthe number of cells sufficient for measurement of all measurement itemsby adding the sample, the sample is concentrated by the centrifugalseparation unit 38 to newly sample the sample by concentrating thesample in the centrifugal separation unit 38 and mixing a new sample.

Next, in S20 of FIG. 11, the control unit 4 determines whetherconcentration adjustment of the sample is necessary. Whether theconcentration adjustment of the sample is necessary is determinedaccording to the concentration information of the measurement targetcells in the sample generated in S16 and the particle detection reagentused for preparation of the measurement sample, and concentrationadjustment is deemed necessary when the concentration of the measurementtarget cells in the sample is larger than a predetermined value orsmaller than a predetermined value. On the other hand, when theconcentration of the measurement target cells in the sample is equal toa predetermined value, it is determined that concentration adjustment isunnecessary. The predetermined value may be a fixed value or may have acertain range.

In the example of FIG. 13A, for example, since the number of white bloodcells necessary for measurement of regulatory T cells is 1×10⁴ cells andthe amount of sample is 10 the leukocyte concentration as themeasurement target cells in the sample of the above-mentionedpredetermined value becomes 1×10³ cells/μl. Here, as shown in FIG. 14,concentration adjustment is deemed not required as a result of thepre-measurement of the sample, since the leukocyte concentration ofsample A is 1×10³/Ml, the leukocyte concentration of sample B is1×10⁴/Ml, the leukocyte concentration of sample C is 1×10²/Ml. On theother hand, in sample B the leukocyte concentration is larger than thepredetermined value, and in sample C the leukocyte concentration issmaller than the predetermined value, so it is determined thatconcentration adjustment is necessary.

Note that the proportion of measurement target cells in the sample maybe low depending on the measurement item. For example, this example is ahematopoietic stem cell (measurement target cell) in the bone marrow. Inthis case, whether concentration adjustment of the sample is necessaryis determined according to the information of the ratio of themeasurement target cells contained in a certain fixed amount of samplecontained in the cell detection reagent. Concentration adjustment of thesample in this case may be performed by centrifugal separation to bedescribed later. When the flow cytometer is provided with a sortingfunction, the measurement target cells also may be collected andconcentrated from the sample using the sorting function.

When it is determined in S20 of FIG. 11 that concentration adjustment ofthe sample is unnecessary, the control unit 4 proceeds to S21 andcontrols the sample dispensing unit 30A to suction a predeterminedsample volume by the nozzle 300, and discharge the suctioned sample intothe dispensing tube 11. Then, the control unit 4 controls the tubetransfer unit 36 to set the dispensing tube 11 holding the dispensedsample in the centrifugal separation unit 38 of the sample preparationunit 3, and then in S23, the reagent dispensing unit 30B is controlledto dispense cell detection reagent and reagent other than the celldetection reagent to the dispensing tube 11 in order to prepare themeasurement based on information relating to preparation of themeasurement sample.

Specifically, the control unit 4 controls the operations of samplepreparation (dispensing, diluting, washing and the like) by the samplepreparation unit 3 based on the type of the cell detection reagentnecessary for preparing the measurement sample, the type of the reagentother than the cell detection reagent necessary for preparation of themeasurement sample, the order of dispensing each reagent, the dispensingamount of each reagent, the amount of sample and the like included inthe information related to preparation of the measurement sample Uponpreparation of the measurement sample, the control unit 4 also maycontrol the centrifugal separation unit 38 as necessary to performcentrifugal separation, and control the reagent dispensing unit 30B soas to remove the supernatant in the dispensing tube 11. Informationrelating to the temperature at the time of dispensing each reagent alsomay be included in the information on the preparation of the measurementsample, and the control unit 4 controls the temperature adjustment unit398, convection generating units 399 a and 399 b to adjust thetemperature of the centrifugal separation unit 38 when dispensing eachreagent based on the information relating to the temperature at the timeof dispensing each reagent included in the information on thepreparation of the measurement sample.

For example, when describing sample A in FIG. 14 as an example, thecontrol section 4 controls the sample dispensing unit 30A to dispensepart of sample A from container 10 to another dispensing tube 11 that isdifferent from the dispensing tube 11 used in the premeasurement asshown in FIG. 15 (a). The amount of sample to be dispensed into thedispensing tube 11 satisfies the required number of cells if 10 μl ofsample A is dispensed into the dispensing tube 11 for preparation of themeasurement sample since the number of white blood cells necessary formeasurement of regulatory T cells is 1×10⁴ as shown in FIG. 13A, and theleukocyte concentration of sample A is 10³/Ml.

Next, the control unit 4 controls the tube transfer unit 36 to transferand set the dispensing tube 11 containing sample A to the centrifugalseparation unit 38 of the sample preparation unit 3. When analyzingregulatory T cells using peripheral blood as a sample in accordance withinformation relating to the preparation of the measurement sample shownin FIG. 13A, it is preferable to first hemolyze the erythrocytes, sothat the control unit 4 performs reagent dispensing of 5 μl of hemolyticagent set in the reagent setting unit 35 into the dispensing tube 11containing sample A under the control of the unit 30B. Subsequently, thecontrol unit 4 controls the reagent dispensing unit 30B to dispense 10μl of the cell detection reagent 2 set in the reagent setting unit 35,and 5 μl of cell detection reagent 3 into dispensing tube 11 containingsample according to the information relating to preparation of themeasurement sample shown in FIG. 13A. Subsequently, the control unit 4controls the reagent dispensing unit 30B to dispense 10 μl of cellmembrane penetrating agent is set in the reagent setting unit 35 in thedispensing tube 11 containing the cells reacted with the cell detectionreagent 2 and the cell detection reagent 3. Then, the control unit 4controls the reagent dispensing unit 30B to dispense 10 μl of the celldetection reagent 4 set in the reagent setting unit 35 to the dispensingtube 11 in which 10 μl of the cell membrane penetrating agent has beenmixed. Finally, the control unit 4 controls the reagent dispensing unit30B to dispense 5 μl of the cell detection reagent 1 set in the reagentsetting unit 35 to the dispensing tube 11 in which cell detectionreagent 4 was mixed. Note that the cell detection reagent 1 is a nuclearstain.

In the example of FIG. 15 (a), it is preferable to incubate for 15 to 30minutes in order to sufficiently react the cells with the antibody aftereach cell detection reagent is dispensed. After the incubation, thecells also may be washed before dispensing the next cell detectionreagent. The washing method is not limited; for example, centrifugalseparation of the dispensing tube 11 containing the cells may beperformed by the centrifugal separation unit 38 to remove of thesupernatant, and diluting liquid or the like may be added to resuspendthe cells.

Then, in S24, the control unit 4 transfers the dispensing tube 11containing the measurement sample prepared as described above to themeasurement unit 2 under the control of the tube transfer unit 36. Whenthe dispensing tube 11 is transported to a predetermined position 29(shown in FIG. 3) of the measurement unit 2, the measurement sample isdischarged from the dispensing tube 11 by the suction unit 29 a that canmove up and down in the vertical direction (Z direction), and the samplesupplied to the flow cell 20 of the measurement unit 2. In this way thecontrol unit 4 performs the main measurement of the measurement sampleby the flow cytometry method using the measurement unit 2 in S25.

On the other hand, in S20, when the control unit 4 determines thatconcentration adjustment of the sample is necessary, the control unit 4proceeds to S22 and adjusts the concentration of the sample in thesample container 10. Specifically, when the concentration of themeasurement target cells in the sample is larger than the predeterminedvalue, the control unit 4 can make the concentration of the measurementtarget cells in the sample a predetermined value by diluting the sample.

Referring to the example of sample B in FIG. 14, the control unit 4controls the sample dispensing unit 30A so that the sample dispensingtube 11 dispenses part of the sample B from the sample container 10, asshown in FIG. 15 (b). The amount of sample dispensed into the dispensingtube 11 can satisfy the required number of cells if 1 μl of sample B isdispensed into dispensing tube 11 for preparing a measurement samplesince the leukocyte count required for measurement of regulatory T cellsis 1×10⁴ and the leukocyte concentration of sample B is 1×10⁴/Ml, asshown in FIG. 13A. Then, the leukocyte concentration of sample B can beset as the predetermined value by diluting 10-fold with diluting liquidto make the amount of sample B 10 μl since the leukocyte concentrationin sample B is 10-fold higher than the predetermined value of 1×103cells/μl.

The control unit 4 also concentrates the sample when the concentrationof the measurement target cells in the sample is smaller than apredetermined value. Concentration of the sample can be performed, forexample, by centrifugal separation by the centrifugal separation unit38. The concentration of the measurement target cells in the sample canbe set to a predetermined value by concentrating the sample.

Referring sample C in FIG. 14 as an example, the control unit 4 controlsthe sample dispensing unit 30A to dispense part of sample C from thesample container 10 to another dispensing tube 11 for pre-measurement,as shown in FIG. 15 (c). The amount of sample to be dispensed into thedispensing tube 11 satisfies the required number of cells if 100 μl ofsample B is dispensed into the dispensing tube 11 for preparation of themeasurement sample since the number of leukocytes necessary formeasurement of regulatory T cells is 1×10⁴ cells, and the leukocyteconcentration of sample C is 1×10²/μl. Then, the control unit 4 controlsthe centrifugal separation unit 38 to centrifugally separate sample Cand the reagent dispensing unit 30B to suction and remove thesupernatant from the dispensing tube 11, and mix the dilution liquid andsample to 10 μl to set the leukocyte concentration to a predeterminedvalue. Note that in this case it is preferable to concentrate red bloodcells after hemolysis.

When the concentration adjustment of the sample is completed the processcontinues to S23 of FIG. 11, and the reagent dispensing unit 30B iscontrolled based on the information relating to the preparation of themeasurement sample, and cell detection reagent and reagent other thanthe cell detection reagent are dispensed into the dispensing tube 11 toprepare a measurement sample. Then, in S24, the control unit 4 controlsthe tube transfer unit 36 to transport the dispensing tube 11 containingthe prepared measurement sample to the measurement unit 2, and in S25 InS25, the main measurement of the measurement sample is performed by themeasurement unit 2 by flow cytometry.

Then, in S26, the control unit 4 transmits the measurement data of themain measurement from the measurement unit 2 to the analysis unit 5. Theprocessing unit 50 of the analysis unit 5 analyzes the sample using themeasurement data of this measurement, and determines whether there is anabnormality in the measurement target cells in the sample.

Second Embodiment

A second embodiment of the operation of the control unit 4 will bedescribed with reference to FIG. 11 and FIG. 12B. Note that thefollowing flow is merely an example, and the embodiment is not limitedto the following flow. The second embodiment is an example in whichmeasurement samples for measuring each measurement item of apredetermined test item are prepared with different dispensing tubes 11.FIG. 13B is an example of information related to the preparation of themeasurement sample recorded in the storage unit 42 of the control unit 4in the second embodiment. In the second embodiment, four kinds ofmeasurement samples (measurement samples 1 to 4) are prepared by mixingcell detection reagents 2 to 5 as cell detection reagents with samplesby separate dispensing tubes 11, respectively, and the operation in thiscase will be described. Note that the same operation is performed whenpreparing measurement samples other than three types.

First, S10 to S18 of FIG. 11 are the same as those of the firstembodiment, so detailed description thereof will be omitted here.

Next, the control unit determines measurement items in S19. In S190 ofFIG. 12B, the number of measurement target cells in the sample in thesample container 10 (total number of cells) is calculated based on theconcentration information of the measurement target cells in the samplegenerated in S16 and the sample amount in the sample container 10. Next,in step S191, the control unit 4 determines whether the calculatednumber of measurement target cells in the sample is equal to or largerthan the number of cells sufficient for measurement of all themeasurement items of the test item. The number of cells necessary formeasurement of this measurement item is obtained from informationrelating to preparation of the measurement sample.

In the example of FIG. 13B, since the leukocyte count necessary formeasurement of regulatory T cells is 5×10³ cells in the measurement itemusing the cell detection reagent 2, 1×10³ cells in the measurement itemusing the cell detection reagent 3, 1×10³ cells in the measurement itemusing the cell detection reagent 4, and 10×10³ cells in the measurementitem using the cell detection reagent 5, a total of 17×10³ cells arerequired. As shown in FIG. 14, as a result of the pre-measurement of thesample it is known that the leukocyte concentration of sample A is1×10³/Ml, the sample volume is 1 ml, and the number of leukocytemeasurement target cells in the sample A is 1×10⁶, so sample A satisfiesthe measurement conditions of all the measurement items. The number ofleukocyte measurement target cells in the sample B is 1×10⁷ cells and1×10⁵ cells, respectively, so that the samples B and C satisfy themeasurement conditions of all the measurement items. Therefore, S191becomes “YES” and the flow advances to S192, and the control unit 4determines the measurement items of all the measurement items. Thedetermination result also is recorded in the storage unit 42 andtransmitted to the analysis unit 5, and the process proceeds to S20 ofFIG. 11.

On the other hand, although not shown in the figure, the number ofleukocyte measurement target cells in the sample will not satisfy themeasurement conditions of all measurement items when, for example, thenumber of leukocyte measurement target cells is less than 16×10³ cells.In this case, in S191, the control unit 4 determines that the number ofmeasurement target cells in the sample is not a sufficient amount formeasurement of all the measurement items, and the process proceeds toS193.

Next, in step S191, the control unit 4 determines whether the number ofmeasurement target cells in the sample is equal to or larger than thenumber of cells sufficient for measurement of all the measurement itemsof the test item. In the example of FIG. 13B, when the number ofleukocyte measurement target cells in the sample is, for example, 16×10³cells, the determination is “YES” in S193, the process proceeds to S194,and the control unit 4 determines the measurement items to be measuredbased on the number of measurement target cells in the sample and theorder of priority of the measurement items that are given priority whenthe number of cells included in the information on preparation of themeasurement sample is insufficient. That is, it is determined whetherthe number of cells satisfying the measurement condition of themeasurement item can be ensured in descending order of priority, and themeasurement item that can ensure the number of cells satisfies themeasurement condition is determined as the measurement item. Thepriority order for determining this measurement item is obtained frominformation relating to the preparation of the measurement sample.

In the example of FIG. 13B, when the number of leukocyte measurementtarget cells in the sample is, for example, 16×10³ cells, themeasurement item using the cell detection reagent 5 having the firstpriority is determined first. The number of cells necessary formeasuring the first measurement item is 10×10³ cells, so that themeasurement condition of the measurement item is satisfied. Next, themeasurement item using the cell detection reagent 2 having the secondhighest priority is determined; since the number of cells necessary forthe measurement of the second measurement item is 5×10³ cells, and theremaining number of leukocyte measurement target cells in the sample is6×10³, the measurement condition of the measurement item is satisfied.Next, the measurement item using the cell detection reagent 4 with thethird priority is determined; since the number of cells necessary forthe measurement of the third measurement item is 1×10³ cells, and theremaining number of leukocyte measurement target cells in the sample is1×10³, the measurement condition of the measurement item is satisfied.Finally, the measurement item using the cell detection reagent 3 withthe fourth priority is determined; since the number of cells necessaryfor the measurement of the fourth measurement item is 1×10³ cells, andthe number of remaining white blood cells in the sample in the sample is0, the measurement condition of the measurement item is not satisfied.Therefore, in this case, the control unit 4 determines the measurementitems using the cell detection reagents 2, 4, and 5 as measurementitems. The determination result also is recorded in the storage unit 42and transmitted to the analysis unit 5, and the process proceeds to S20of FIG. 11.

On the other hand, if it is determined in S193 that the number ofmeasurement target cells in the sample is not sufficient for allmeasurement items, the control unit 4 proceeds to S195 and determinesthe number of cells necessary for measuring the measurement item is notensured and notifies the analyzing unit 5 that a measurement sample cannot be prepared.

Next, in S20 of FIG. 11, the control unit 4 determines whetherconcentration adjustment of the sample is necessary. In the secondembodiment as shown in the example of FIG. 13B, the concentration of themeasurement target cells (leukocytes) in the sample used for thepreparation is different for each measurement item; the predeterminedvalue of the leukocyte concentration of the sample required for themeasurement item using cell detection reagent 2 is 5×10² cells/μl, thepredetermined value of the leukocyte concentration of the samplerequired for the measurement item using the cell detection reagent 3 is1×10² cells/μl, the leukocyte concentration of the sample required forthe measurement item using the cell detection reagent 4 is 1×10²cells/μl, and the predetermined value of the leukocyte concentration ofthe sample required for the measurement item using the cell detectionreagent 5 is 1×10³ cells/μl. Therefore, whether concentration adjustmentis necessary based on the concentration information of the measurementtarget cells in the sample is determined for each measurement itemgenerated in S16, and when the concentration of the measurement targetcells in the sample is larger than the predetermined value or smallerthan the predetermined value, the control unit 4 determines thatconcentration adjustment is necessary, and the process continues to S22.Note that since the method of S22 concentration adjustment is the sameas that of the first embodiment, a detailed description thereof will beomitted. On the other hand, when the concentration of the measurementtarget cell in the sample is the predetermined value, the control unit 4determines that concentration adjustment is unnecessary, and proceeds toS21.

In S21, the control unit 4 controls the sample dispensing unit 30A tosuction a predetermined sample amount of sample for each measurementitem by the nozzle 300, and discharges the sample into each dispensingtube 11. Then, the control unit 4 controls the tube transfer unit 36 toset the dispensing tubes 11, into which the samples are dispensed, inthe centrifugal separation unit 38 of the sample preparation unit 3, andin S23 controls the reagent dispensing unit 30B based on the informationrelating to preparation of the measurement sample, and discharges celldetection reagent and reagent other than cell detection reagent into thedispensing tubes 11 for each measurement item to prepare the measurementsample.

In the case of preparing a measurement sample by mixing a plurality ofcell detection reagents with samples by separate dispensing tubes 11 asin the second embodiment, the control unit 4 controls the sampledispensing unit 30A to dispense sample to each dispensing tube 11 toattain the required number of cells corresponding to the cell detectionreagents 2 to 5, respectively. Next, the control unit 4 controls thetube transfer unit 36 to transfer and set each of the dispensing tubes11 containing the sample in the centrifugal separation unit 38 of thesample preparation unit 3. Subsequently, the control unit 4 controls thereagent dispensing unit 30B to dispense 5 μl of the hemolytic agent setin the reagent setting unit 35 to each dispensing tube 11 containing thesample. In addition to the hemolytic agent, 10 μl of the cellpenetrating agent set in the reagent setting section 35 is alsodispensed into the dispensing tube to which the cell detection reagent 5was dispensed under the control of the reagent dispensing unit 30B.Subsequently, the control unit 4 controls the reagent dispensing unit30B to dispense cell detection reagent 2 to 5 set in the reagent settingunit 35 to each dispensing tube 11 containing the sample. The controlunit 4 also controls the reagent dispensing unit 30B to dispense thenuclear staining solution (cell detection reagent 1) set in the reagentsetting unit 35 to all the dispensing tubes.

Then, in S24, the control unit 4 transfers all the dispensing tubes 11containing the measurement samples prepared as described above to themeasurement unit 2 under the control of the tube transfer unit 36. Inthis way the control unit 4 performs the main measurement of themeasurement samples by the flow cytometry method using the measurementunit 2 in S25.

As described above, according to the sample preparation apparatus 1, thesample preparation method, and the particle analyzer 100 of the aboveembodiments, pre-measurement is performed on the sample, concentrationinformation of the measurement target cells in the sample is generatedbased on the measurement data, and the concentration of the measurementtarget particles in the sample is adjusted according to the generatedconcentration information and the cell detection reagent used forpreparing the measurement sample based on this measurement, and thesample preparation unit 3 prepares the measurement sample. Therefore, itis possible to efficiently prepare a measurement sample including themeasurement target cells at a concentration suitable for the celldetection reagent, and it is possible to analyze the measurement targetcells in the sample with high accuracy.

The operation conditions (the amount of the sample, the type of the celldetection reagent and other reagents necessary for preparation of themeasurement sample, the dispensing order, the dispensing amount and thelike) are controlled based on the information related to the preparationof the measurement sample used for preparing the measurement sample.Therefore, preparation of the measurement sample can be automated.

Other Modifications

Although the embodiment of the particle analyzer has been describedabove, the present invention is not limited to the above-describedembodiment, and various modifications are possible without departingfrom the spirit of the present invention.

For example, although the measurement unit 2 has a function ofpre-measuring a sample containing particles before sample preparationand detecting the measurement target particles contained in the sample,and a function of performing a main measurement of the measurementsample and detecting information related to characteristics of themeasurement target cells for cell analysis by the analyzer 5 in theabove embodiment, the function of pre-measuring the sample alone alsomay be provided. In this case, in the particle analyzer 1, a mainmeasurement unit (second measurement unit) such as a flow cytometer orthe like for performing a main measurement of the measurement sample,and a measurement unit 2 (first measurement unit) for measuring a sampleand detecting measurement target particles in the sample may be providedin particle analyzer 1. Although the second measuring unit may beincluded in the sample preparation apparatus 1, or may be providedseparately from the sample preparation apparatus 1, in which case, aflow cytometer configured as shown in FIG. 2 also may be used as themeasurement unit 2, since the measuring unit 2 is capable of outputtinga signal for detecting the measurement target particles contained in thesample, a light receiving element for acquiring the forward scatteredlight may be provided without a light receiving element for acquiringthe side scattered light and side fluorescent light inasmuch asacquiring the forward scattered light is sufficient. The measurementunit 2 also is not limited to optical particle measurement such as aflow cytometer, and may be an electric resistance type particlemeasurement, a precipitation type particle measurement.

In the above-described embodiment, the centrifugal separation unit 38 ofthe sample preparation unit 3 functions as a concentration adjustmentunit, but the specific configuration for concentration adjustment is notparticularly limited, and a filter, a cell sorter, or the like also maybe used.

Although the sample preparation unit 3 includes the centrifugalseparation unit 38, and the measurement sample is prepared by thecentrifugal separation unit 38 in the above embodiment, the specificconfiguration for preparing the measurement sample is not particularlylimited inasmuch as the measurement sample can be preparedautomatically.

In the above embodiment, The sample test items and measurement items aredecided based on information relating to the cell detection reagent setin the reagent setting unit 35 by the receiving unit 39 installed in thereagent setting unit 35 of the sample preparation unit 3. However,information specifying measurement items also may be acquired byproviding a receiving unit such as a barcode reader or RFID reader orthe like capable of reading a barcode or the like in the samplecontainer setting unit 31 and affixing a barcode, tag or the likecontaining information specifying measurement items on the samplecontainer 10, so that information specifying the measurement item can beacquired by reading the barcode or the like attached to the samplecontainer 10. The information for specifying the measurement item is notlimited insofar as it can specify the measurement item, and may be, forexample, the name of the measurement item, identification information orname of the reagent necessary for measuring the measurement item and thelike.

In the above-described embodiments, the control unit 4 also maydetermine the number of measurement samples to be prepared by the samplepreparation unit 3 based on the type of the particle detection reagentused for preparing the measurement sample and the number of measurementtarget particles in the sample in the sample container. For example,information relating to the number of measurement samples (dispensingtubes 11) required for a measurement can be included in the informationrelating to the preparation of measurement sample, and the control unit4 may control the measurement sample preparation unit 3 to prepare apredetermined number of measurement samples based on information relatedto the number of measurement target cells in the sample in the samplecontainer 10 and the number of measurement samples (dispensing tubes 11)required for the measurement included in the information relating to thepreparation of the measurement sample.

In the above-described embodiment, the sample preparation apparatus 1 isconfigured as one device integrally incorporating the measurement unit2, the sample preparation unit 3, and the control unit 4. However, inaddition to the above embodiments, the invention also may be a samplepreparation system 1′ in which the measurement unit 2, sample preparingunit 3 and control unit 4 are independent devices configured asmeasurement device 2′, sample preparation device 3′ and control device4′, such that the measurement device 2′, sample preparation device 3′,and analyzer 5′ are the sample preparation system connected to thecontrol device 4. Note that the measurement device 2′, the samplepreparation device 3′, the control device 4′, and the analyzer 5′ havethe same or substantially the same configuration as the measurement unit2, the sample preparation unit 3, the control unit 4 and the analyzer 5in FIG. 1. In this case, the signal processing unit 6 in FIG. 1 isincluded in the measurement device 2′, and the communication I/F isincluded in each of the devices 2′ to 5′.

What is claimed is:
 1. A sample preparation apparatus comprising: a measurement unit configured to measure a sample that includes particles acquired from a sample container and to detect measurement target particles in the sample; a sample preparation unit configured to adjust a concentration of the measurement target particles in the sample and to mix the sample with a plurality of types of particle detection reagents that include a particle labeling substance, and to prepare a measurement sample; a control unit configured to control the sample preparation unit so as to generate concentration information of the measurement target particles in the sample based on measurement data from the measurement unit, and to adjust the concentration of the measurement target particles in the sample according to the generated concentration information and the plurality of types of the particle detection reagents mixed with the measurement sample.
 2. The sample preparation apparatus of claim 1, wherein the control unit controls the sample preparation unit so as to mix the sample whose concentration of the measurement target particles has been adjusted with the particle detection reagents by an amount corresponding to the types of the particle detection reagents used for preparation of the measurement sample.
 3. The sample preparation apparatus of claim 1, wherein the control unit controls the sample preparation unit so as to adjust the concentration of the measurement target particles in the sample by concentrating or diluting the sample obtained from the sample container.
 4. The sample preparation apparatus according to claim 1, wherein the sample preparation unit comprises a concentration adjusting unit configured to adjust the concentration of the measuring target particles in the sample acquired from the sample container, and a reagent dispensing unit configured to dispense the particle detection reagents to the sample for which the concentration of the measurement target particles has been adjusted.
 5. The sample preparation apparatus according to claim 4, wherein the concentration adjusting unit includes a centrifugal separation unit or a cell sorter.
 6. The sample preparation apparatus according to claim 1, wherein the measurement unit comprises a flow cell, a light source that irradiates light on the sample passing through the flow cell, and a light receiving element that detects optical information from the measurement target particles in the sample and converts the optical information into an electric signal.
 7. The sample preparation apparatus according to claim 6, wherein the measurement unit is supplied with the measurement sample prepared in the sample preparation unit, and performs measurements on the measurement sample.
 8. The sample preparation apparatus according to claim 1, wherein the sample preparation unit comprises a sample dispensing unit configured to suction the sample from the sample container; the measurement unit measures a part of the sample in the sample container suctioned by the sample dispensing unit; and the sample preparation unit prepares the measurement sample at least from another part of the sample in the sample container suctioned by the sample dispensing unit.
 9. The sample preparation apparatus according to claim 1, further comprising: a storage unit that stores information on the preparation of the measurement sample; and the control unit is configured to read the information corresponding to the particle detection reagents to be used in the preparation of the measurement sample, and control the sample preparation unit to prepare the measurement sample based on the read information.
 10. The sample preparation apparatus according to claim 9, wherein the information on the preparation of the measurement sample comprises at least an amount of the respective particle detection reagents required to prepare the measurement sample, an amount of the sample mixed with the particle detection reagent, and a number of the measurement target particles in the sample mixed with the particle detection reagents.
 11. The sample preparation apparatus according to claim 9, wherein the sample preparation unit prepares the measurement sample from the sample whose concentration of the measurement target particles has been adjusted and the plurality of types of particle detection reagents; the information on the preparation of the measurement sample stored in the storage unit comprises an order of adding the plurality of types of particle detection reagents to the sample; and the control unit controls the sample preparation unit so as to mix the plurality of types of particle detection reagents to be used for the preparation of the measurement sample into the sample in the order based on the information on the preparation of the measurement sample read from the storage unit.
 12. The sample preparation apparatus according to claim 1, wherein the control unit is configured to calculate a number of measurement target particles in the sample in the sample container from a liquid amount of the sample in the sample container and the generated concentration information; and the control unit determines a number of measurement samples to be prepared by the sample preparation unit based on the types of particle detection reagents and the calculated number of measurement target particles in the sample in the sample container.
 13. The sample preparation apparatus according to claim 1, further comprising: a reception unit that receives an input of identification information of a particle detection reagents used for preparing the measurement sample.
 14. The sample preparation apparatus according to claim 1, wherein the control unit determines the types of particle detection reagents to be used for the preparation of the measurement sample based on the identification information received by the reception unit.
 15. The sample preparation apparatus according to claim 1, wherein the control unit determines the types of particle detection reagents to be used for preparing the measurement sample based on a measurement item of the sample in the sample container.
 16. The sample preparation apparatus according to claim 15, wherein the control unit determines the measurement item based on the generated concentration information.
 17. The sample preparation device according to claim 1, wherein the particles are cells.
 18. The sample preparation device according to claim 1, wherein the plurality of types of particle detection reagents comprises a first particle detection reagent and a second particle detection reagent, the first particle detection reagent comprises a first fluorescent substance whose excitation wavelength is a first wavelength, and the second particle detection reagent comprises a second fluorescent substance whose excitation wavelength is a second wavelength which is different from the first wavelength.
 19. A sample preparation method, comprising: generating concentration information of measurement target particles in a sample: acquiring type information of a plurality of particle detection reagents used for preparing a measurement sample; adjusting a concentration of measurement target particles in the sample used in the preparation of the measurement sample according to the generated concentration information and the acquired type information; and preparing the measurement sample by mixing the sample whose concentration of the measurement target particles is adjusted with the plurality of the particle detection reagents.
 20. A sample preparation system comprising: a measuring device that measures a sample containing particles acquired from a sample container and detects measurement target particles in the sample; a sample preparation apparatus that adjusts a concentration of the measurement target particles in the sample acquired from the sample container and mixes the sample with a plurality of types of particle detection reagents to prepare a measurement sample; a control device that is connected to the measuring device and the sample preparation apparatus, that controls the sample preparation apparatus so as to generate concentration information of the measurement target particles in the sample in the sample container based on measurement data of the measuring device, and that adjusts the concentration of the measurement target particles in the sample acquired from the sample container according to the generated concentration information and the plurality of types of the particle detection reagents used for preparing the measurement sample. 